HPLC-MS/MS同时测定感冒安颗粒中5种黄酮成分的含量

Simultaneous determination of five flavonoids in Ganmao’an granules by HPLC-MS/MS

目的建立HPLC-MS/MS方法测定感冒安颗粒中5种黄酮成分含量。方法色谱条件:色谱柱为Kromasil C_(18)柱(4.6 mm×150 mm×4.6 mm,5μm,100Å),流动相为甲醇(A)-0.1%甲酸(B),梯度洗脱(0~20 min,35%A;20~40 min,45%A),流速为1.0 ml/min,柱温为25℃;质谱条件:电喷雾电离源(ESI源)负离子扫描,多反应监测(MRM)模式检测。5种黄酮检测的离子对:芦丁m/z 609.1→300.1、金丝桃苷和异槲皮苷m/z 463.0→300.1、槲皮素m/z 301.0→151.0、木犀草素m/z 285.0→132.9。结果5个成分在一定浓度范围内与峰面积具有良好的线性关系,相关系数r均>0.9991;平均加样回收率在100.63%~102.81%之间,RSD范围在0.67%~2.07%。10批感冒安颗粒中芦丁、金丝桃苷、异槲皮苷、槲皮素和木犀草素的含量测定结果分别为32.23~479.83、0.291~1.825、11.44~20.54、6.32~18.41、3.46~6.51μg/g。结论建立的HPLCMS/MS法灵敏度高、专属性强,适合同时测定感冒安颗粒中5种黄酮类成分的含量。

Objective To develop a high performance liquid chromatography-tandem mass spectrometry(HPLCMS/MS)method for simultaneous determination of five flavonoids in Ganmao’an granules(GMA).Methods Chromatographic separation was achieved on a Kromasil C_(18) Column(150 mm×4.6 mm,5μm,100Å),which was eluted with methanol(A)-0.1%formic acid(B)at the flow rate of 1.0 ml/min.The gradient condition was as follows:0-20 min,35%A,and 20-40 min,45%A.The column temperature was 25°C.Analytes were detected using a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization source in the negative ion scanning.The multiple reaction monitoring mode was used for qualitative analysis.For each flavonoid,two precursor ion/product ion transitions were chosen:lutin m/z 609.1→300.1,hyperin and isoquercitrin m/z 463.0→300.1,quercetin m/z 301.0→151.0,luteolin m/z 285.0→132.9.Results Five flavonoids showed the good relationships within their own concentration ranges(correlation coefficient r>0.9991),whose average recoveries were in the range of 100.63%-102.81%with RSDs of 0.67%-2.07%.The content results of rutin,hyperoside,isoquercetin,quercetin,and luteolin in 10 batches of GMA were 32.23-479.83,0.291-1.825,11.44-20.54,6.32-18.41,3.46-6.51μg/g,respectively.Conclusion The results indicated that the developed method was sensitive,accurate and could provide excellent specificity for simultaneous determination of five flavonoids in GMA.